Cytotoxic Evaluation in HaCaT Cells of the Pa.7 Bacteriophage from Cutibacterium (Propionibacterium) acnes, Free and Encapsulated Within Liposomes.

Introduction: Acne is a multifactorial disease involving the colonization of skin follicles by Cutibacterium (formerly Propionibacterium) acnes. A combination of different retinoid-derived products, antibiotics, and hormonal antiandrogens are used to treat the disease, but these treatments require extended periods, may have secondary effects, are expensive, and not always effective. Owing to antibiotic resistance, the use of bacteriophages has been proposed as an alternative treatment. However, if they are intended for a cosmetic or pharmaceutical use, it is necessary to evaluate the safety of the phages and the preparations containing them. Materials and Methods: In this study, the cytotoxicity of Pa.7 bacteriophage was evaluated in HaCaT cells, along with a liposome suitable for their encapsulation, using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and trypan blue assays. Results: We found that Pa.7 was not cytotoxic for HaCaT cells. Also, 30 mM of liposomes, or below are considered noncytotoxic concentrations. Conclusion: Phages encapsulated in the liposomes presented in this study can be used safely for skin treatments.

Bioremediation of a Sewage-Contaminated Tropical Swamp Through Bioaugmentation with a Microalgae-Predominant Microbial Consortium.

Bioaugmentation has many applications as a bioremediation technique. It is usually performed by bacteria but microalgal consortia also have great potential for bioremediation. This study evaluated the ability of a microbial consortium with predominance of microalgae (MCPM) to decontaminate the water of the Mallorquín tropical swamp. The Mallorquín Swamp is a natural water reservoir, essential for the ecological and hydric balance of the region, as well as for the fishermen, but has received sewage for more than 20 years. Microalgae for bioremediation purposes were isolated from the swamp, selected, cultured in bioreactors, and poured back into the Mallorquín waters. After bioaugmentation, there was a significant reduction in the BOD5 (98%), nitrates (58%), enterococci (92%), and total coliforms (100%). Notably, despite the MCPM bioaugmentation, the original richness and abundance of phytoplankton in the Mallorquín swamp was not disrupted. These results confirm the benefits of phycoremediation as an effective tool for on-site bioremediation of natural water bodies and show an effective phycoremediation at a large scale without altering the autochthonous microalgae community. This constitutes the first report of a successful MCPM intervention performed at this scale in a natural swamp in Colombia. Supplementary Information: The online version contains supplementary material available at 10.1007/s12088-021-00990-y.

Phages in Anaerobic Systems.

Since the discovery of phages in 1915, these viruses have been studied mostly in aerobic systems, or without considering the availability of oxygen as a variable that may affect the interaction between the virus and its host. However, with such great abundance of anaerobic environments on the planet, the effect that a lack of oxygen can have on the phage-bacteria relationship is an important consideration. There are few studies on obligate anaerobes that investigate the role of anoxia in causing infection. In the case of facultative anaerobes, it is a well-known fact that their shifting from an aerobic environment to an anaerobic one involves metabolic changes in the bacteria. As the phage infection process depends on the metabolic state of the host bacteria, these changes are also expected to affect the phage infection cycle. This review summarizes the available information on phages active on facultative and obligate anaerobes and discusses how anaerobiosis can be an important parameter in phage infection, especially among facultative anaerobes.

Paving the Way to Unveil the Diversity and Evolution of Phage Genomes.

Phage biology has been developing for the last hundred years, and the potential of phages as tools and treatments has been known since their early discovery. However, the lack of knowledge of the molecular mechanisms coded in phage genomes hindered the development of the field. With current molecular methods, the last decade has been a resurgence of the field. The Special Issue on "Diversity and Evolution of Phage Genomes" is a great example with its 17 manuscripts published. It covers some of the latest methods to sample and characterize environmental and host associated viromes, considering experimental biases and computational developments. Furthermore, the use of molecular tools coupled with traditional methods has allowed to isolate and characterize viruses from different hosts and environments with such diversity that even a new viral class is being proposed. The viruses described cover all different phage families and lifestyles. However, is not only about diversity; the molecular evolution is studied in a set of manuscripts looking at phage-host interactions and their capacity to uncover the frequency and type of mutations behind the bacterial resistance mechanisms and viral pathogenesis, and such methods are opening new ways into identifying potential receptors and characterizing the bacterial host range.

Host Resistance, Genomics and Population Dynamics in a Salmonella Enteritidis and Phage System.

Bacteriophages represent an alternative solution to control bacterial infections. When interacting, bacteria and phage can evolve, and this relationship is described as antagonistic coevolution, a pattern that does not fit all models. In this work, the model consisted of a microcosm of Salmonella enterica serovar Enteritidis and φSan23 phage. Samples were taken for 12 days every 48 h. Bacteria and phage samples were collected; and isolated bacteria from each time point were challenged against phages from previous, contemporary, and subsequent time points. The phage plaque tests, with the genomics analyses, showed a mutational asymmetry dynamic in favor of the bacteria instead of antagonistic coevolution. This is important for future phage-therapy applications, so we decided to explore the population dynamics of Salmonella under different conditions: pressure of one phage, a combination of phages, and phages plus an antibiotic. The data from cultures with single and multiple phages, and antibiotics, were used to create a mathematical model exploring population and resistance dynamics of Salmonella under these treatments, suggesting a nonlethal, growth-inhibiting antibiotic may decrease resistance to phage-therapy cocktails. These data provide a deep insight into bacterial dynamics under different conditions and serve as additional criteria to select phages and antibiotics for phage-therapy.

The genomic study of an environmental isolate of Scedosporium apiospermum shows its metabolic potential to degrade hydrocarbons.

Crude oil contamination of soils and waters is a worldwide problem, which has been actively addressed in recent years. Sequencing genomes of microorganisms involved in the degradation of hydrocarbons have allowed the identification of several promoters, genes, and degradation pathways of these contaminants. This knowledge allows a better understanding of the functional dynamics of microbial degradation. Here, we report a first draft of the 44.2 Mbp genome assembly of an environmental strain of the fungus Scedosporium apiospermum. The assembly consisted of 178 high-quality DNA scaffolds with 1.93% of sequence repeats identified. A total of 11,195 protein-coding genes were predicted including a diverse group of gene families involved in hydrocarbon degradation pathways like dioxygenases and cytochrome P450. The metabolic pathways identified in the genome can potentially degrade hydrocarbons like chloroalkane/alkene, chorocyclohexane, and chlorobenzene, benzoate, aminobenzoate, fluorobenzoate, toluene, caprolactam, geraniol, naphthalene, styrene, atrazine, dioxin, xylene, ethylbenzene, and polycyclic aromatic hydrocarbons. The comparison analysis between this strain and the previous sequenced clinical strain showed important differences in terms of annotated genes involved in the hydrocarbon degradation process.

Phage ΦPan70, a Putative Temperate Phage, Controls Pseudomonas aeruginosa in Planktonic, Biofilm and Burn Mouse Model Assays.

Pseudomonas aeruginosa is one of the Multi-Drug-Resistant organisms most frequently isolated worldwide and, because of a shortage of new antibiotics, bacteriophages are considered an alternative for its treatment. Previously, P. aeruginosa phages were isolated and best candidates were chosen based on their ability to form clear plaques and their host range. This work aimed to characterize one of those phages, ΦPan70, preliminarily identified as a good candidate for phage-therapy. We performed infection curves, biofilm removal assays, transmission-electron-microscopy, pulsed-field-gel-electrophoresis, and studied the in vivo ΦPan70 biological activity in the burned mouse model. ΦPan70 was classified as a member of the Myoviridae family and, in both planktonic cells and biofilms, was responsible for a significant reduction in the bacterial population. The burned mouse model showed an animal survival between 80% and 100%, significantly different from the control animals (0%). However, analysis of the ΦPan70 genome revealed that it was 64% identical to F10, a temperate P. aeruginosa phage. Gene annotation indicated ΦPan70 as a new, but possible temperate phage, therefore not ideal for phage-therapy. Based on this, we recommend genome sequence analysis as an early step to select candidate phages for potential application in phage-therapy, before entering into a more intensive characterization.

Positive effects of bacterial diversity on ecosystem functioning driven by complementarity effects in a bioremediation context.

Despite their importance as ecosystem drivers, our understanding of the influence of bacterial diversity on ecosystem functioning is limited. After identifying twelve bacterial strains from two petroleum-contaminated sites, we experimentally explored the impact of biodiversity on total density by manipulating the number of strains in culture. Irrespective of the origin of the bacteria relative to the contaminant, biodiversity positively influenced total density. However, bacteria cultured in the crude oil of their origin (autochthonous) reached higher densities than bacteria from another origin (allochthonous) and the relationship between diversity and density was stronger for autochthonous bacteria. By measuring the relative contribution of each strain to total density we showed that the observed positive effect of increasing diversity on total density was mainly due to positive interactions among species and not the presence of a particular species. Our findings can be explained by the complex chemical composition of crude oil and the necessity of a diverse array of organisms with complementary enzymatic capacities to achieve its degradation. The long term exposure to a contaminant may have allowed different bacteria to become adapted to the use of different fractions of the crude, resulting in higher complementarity in resource use in autochthonous bacteria compared to allochthonous ones. Our results could help improve the success of bioaugmentation as a bioremediation technique by suggesting the use of a diversified set of autochthonous organisms.

Co-evolutionary dynamics of the bacteria Vibrio sp. CV1 and phages V1G, V1P1, and V1P2: implications for phage therapy.

Bacterial infections are the second largest cause of mortality in shrimp hatcheries. Among them, bacteria from the genus Vibrio constitute a major threat. As the use of antibiotics may be ineffective and banned from the food sector, alternatives are required. Historically, phage therapy, which is the use of bacteriophages, is thought to be a promising option to fight against bacterial infections. However, as for antibiotics, resistance can be rapidly developed. Since the emergence of resistance is highly undesirable, a formal characterization of the dynamics of its acquisition is mandatory. Here, we explored the co-evolutionary dynamics of resistance between the bacteria Vibrio sp. CV1 and the phages V1G, V1P1, and V1P2. Single-phage treatments as well as a cocktail composed of the three phages were considered. We found that in the presence of a single phage, bacteria rapidly evolved resistance, and the phages decreased their infectivity, suggesting that monotherapy may be an inefficient treatment to fight against Vibrio infections in shrimp hatcheries. On the contrary, the use of a phage cocktail considerably delayed the evolution of resistance and sustained phage infectivity for periods in which shrimp larvae are most susceptible to bacterial infections, suggesting the simultaneous use of multiple phages as a serious strategy for the control of vibriosis. These findings are very promising in terms of their consequences to different industrial and medical scenarios where bacterial infections are present.

Phylogenetic distance and species richness interactively affect the productivity of bacterial communities.

Our understanding of how biodiversity influences ecosystem functioning is entering a new stage of its development through the incorporation of information about the evolutionary relatedness of species. Bacteria are prime providers of essential ecosystem services, representing an excellent model system to perform biodiversity-ecosystem function research. By using bacteria isolated from petroleum-contaminated sites, we show that communities composed of poorly related species were more productive than those containing highly related species. The nature of the forces controlling this positive effect of phylogenetic diversity on community productivity depended on the number of species in culture. In communities of two species the positive effect of phylogenetic diversity on productivity was driven by changes in the selection effect. Communities of two distantly related species were dominated by the most productive species in monoculture, whereas communities of two closely related species were dominated by the less productive species in monoculture. In communities of four species the positive effect of phylogenetic diversity on productivity was driven by changes in the complementarity effect. In communities composed of four distantly related species the influence of positive interactions such as facilitation, cross-feeding, and niche partitioning seemed to outweigh the influence of negative interactions such as interference. As a consequence the proportion of species favored by the presence of other species increased as they became less related. Multiple facets of biodiversity may influence ecosystem functioning. Here, we present evidence of an interaction between phylogenetic and taxonomic diversity on community productivity, underlining the importance of considering multiple aspects of biodiversity when studying its impact on ecosystem functioning.

The antibacterial activity of phospholipase A(2) type IIA is regulated by the cooperative lipid chain melting behavior in Staphylococcus aureus.

As one of its primary physiological functions, sPLA(2)-IIA appears to act as an antibacterial agent. In particular, sPLA(2)-IIA shows high activity towards Gram-positive bacteria such as Staphylococcus aureus (S. aureus). This antibacterial activity results from the preference of the enzyme towards membranes enriched in anionic lipids, which is a common feature of bacterial membranes. An intriguing aspect observed in a variety of bacterial membranes is the presence of a broad but cooperative lipid chain melting event where the lipids in the membrane transition from a solid-ordered (so) into a liquid-disordered (ld) state close to physiological temperatures. It is known that the enzyme is sensitive to the level of lipid packing, which changes sharply between the so and the ld states. Therefore, it would be expected that the enzyme activity is regulated by the bacterial membrane thermotropic behavior. We determine by FTIR the thermotropic lipid chain melting behavior of S. aureus and find that the activity of sPLA(2)-IIA drops sharply in the so state. The activity of the enzyme is also evaluated in terms of its effects on cell viability, showing that cell survival increases when the bacterial membrane is in the so state during enzyme exposure. These results point to a mechanism by which bacteria can develop increased resistance towards antibacterial agents that act on the membrane through a cooperative increase in the order of the lipid chains. These results show that the physical behavior of the bacterial membrane can play an important role in regulating physiological function in an in vivo system.